The objective of this project is a greater understanding of the structure, biosynthesis, and assembly of influenza virus. The detailed molecular structure of the viral envelope will be further analyzed in biochemical, electron microscopic, and spectroscopic studies. Aberrant glycoproteins which are incorporated into virus particles in the presence of inhibitors of glycosylation will be isolated and their carbohydrate content and biological properties determined. Sulfated macromolecules incorporated into the virion will be further characterized and the cellular site of sulfate incorporation into viral glycoproteins determined. To further analyze the process of viral biosynthesis, we will determine the precise location of viral envelope polypeptides in cytoplasmic and cell surface membranes. The location of the NS polypeptide in the infected cell will also be investigated further. We will examine cells infected with temperature-sensitive mutants by electron microscopy to determine the effects of defects in specific viral functions on the process of replication. The initial steps in replication will be analyzed using autoradiography to determine the fate of input viral RNA. The mechanism of action of two inhibitors of influenza virus replication, adamantanamine hydrochloride and 2-deoxy-2,3-dehydro N- trifluoroacetyl neuraminic acid, will be studied.